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Volume 16, Number 14,
Issue of July 15, 1996
pp. 4344-4359
Copyright ©1996 Society for Neuroscience
[Ca2+]i Elevations Detected by BK
Channels during Ca2+ Influx and Muscarine-Mediated Release
of Ca2+ from Intracellular Stores in Rat Chromaffin
Cells
Received Nov. 7, 1995; revised April 18, 1996; accepted April 23, 1996.
Murali Prakriya,
Christopher R. Solaro, and
Christopher J. Lingle
Washington University School of Medicine, Department of
Anesthesiology, St. Louis, Missouri 63110
Submembrane [Ca2+]i
changes were examined in rat chromaffin cells by monitoring the
activity of an endogenous Ca2+-dependent protein:
the large conductance Ca2+- and voltage-activated
K+ channel (also known as the BK channel). The
Ca2+ and voltage dependence of BK current
inactivation and conductance were calibrated first by using defined
[Ca2+]i salines. This
information was used to examine submembrane
[Ca2+]i elevations
arising out of Ca2+ influx and muscarine-mediated
release of Ca2+ from intracellular stores. During
Ca2+ influx, some BK channels are exposed to
[Ca2+]i of at least 60 µM. However, the distribution of this
[Ca2+]i elevation is
highly nonuniform so that the average
[Ca2+]i detected when all
BK channels are activated is only ~10 µM.
Intracellular dialysis with 1 mM or higher EGTA
spares only the BK channels activated by the highest
[Ca2+]i during influx,
whereas dialysis with 1 mM or higher BAPTA blocks
activation of all BK channels. Submembrane
[Ca2+]i elevations fall
rapidly after termination of short (5 msec) Ca2+
influx steps but persist above 1 µM for several
hundred milliseconds after termination of long (200 msec) influx steps.
In contrast to influx, the submembrane
[Ca2+]i elevations
produced by release of intracellular Ca2+ by
muscarinic actetylcholine receptor (mAChR) activation are much more
uniform and reach peak levels of 3-5 µM. Our
results suggest that during normal action potential activity only
10-20% of BK channels in each chromaffin cell see sufficient
[Ca2+]i to be
activated.
Key words:
BK channels;
calcium;
calcium channels;
calcium
stores;
chromaffin cells;
catecholamine secretion;
K+
channel inactivation
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